title: Crystal structure of Tk-subtilisin folded without propeptide: requirement of propeptide for acceleration of folding.
abstract: Tk-subtilisin (a subtilisin homologue from Thermococcus kodakaraensis) is matured from Pro-Tk-subtilisin upon autoprocessing and degradation of Tk-propeptide. To analyze the folding mechanism of Tk-subtilisin, the crystal structure of the active site mutant of Tk-subtilisin (S324A-subtilisin*), which was refolded in the presence of Ca2+ and absence of Tk-propeptide, was determined at 2.16A resolution. This structure is essentially the same as that of Tk-subtilisin matured from Pro-Tk-subtilisin. S324A-subtilisin* was refolded with a rate constant of 0.17 and 1.8min(-1) at 30 degrees C in the absence and presence of Tk-propeptide, respectively, indicating that Tk-subtilisin does not require Tk-propeptide for folding but requires it for acceleration of folding.
authors.affiliation: Department of Material and Life Science, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: Shun-ichi
authors.initials: S
authors.lastname: Tanaka
authors.forename: Yuki
authors.initials: Y
authors.lastname: Takeuchi
authors.forename: Hiroyoshi
authors.initials: H
authors.lastname: Matsumura
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Koga
authors.forename: Kazufumi
authors.initials: K
authors.lastname: Takano
authors.forename: Shigenori
authors.initials: S
authors.lastname: Kanaya
mesh_terms: D000409:Alanine / Q000737:chemistry / Q000235:genetics; D019843:Archaeal Proteins / Q000737:chemistry* / Q000235:genetics; D002118:Calcium / Q000737:chemistry; D018360:Crystallography, X-Ray; D004792:Enzyme Precursors / Q000737:chemistry* / Q000235:genetics; D010446:Peptide Fragments / Q000737:chemistry* / Q000235:genetics; D011487:Protein Conformation; D017510:Protein Folding; D012694:Serine / Q000737:chemistry / Q000235:genetics; D020860:Subtilisin / Q000737:chemistry* / Q000235:genetics; D013381:Subtilisins / Q000737:chemistry* / Q000235:genetics; D019712:Thermococcus / Q000201:enzymology*
chemical_list: D019843:Archaeal Proteins; D004792:Enzyme Precursors; D010446:Peptide Fragments; C057107:prosubtilisin; D012694:Serine; D013381:Subtilisins; D020860:Subtilisin; D000409:Alanine; D002118:Calcium